Aging induced testicular damage: analyzing the ameliorative potential of Mucuna pruriens seed extract.

Mucuna pruriens Linn. (M. pruriens), a leguminous plant, was used extensively in Ayurveda, to treat male-related infertility. Previous studies have demonstrated antioxidant, androgenic, aphrodisiac, and spermatogenic properties of M. pruriens seed extract. Surprisingly, the biological activities of M. pruriens on aging-induced pathological changes in the testis microenvironment have never been explored and the present study was focused on the testing therapeutic efficacy of M. pruriens on aged rat testis. Male Wistar albino rats were grouped as; adult (3 months), aged (24 months), aged + M. pruriens and adult + M. pruriens (N = 6/group). The extract was administrated at a dose of 200 mg/kg body weight (dosage determined in our previous study) daily by gavage for 60 days. The total and free testosterone, FSH and LH levels were considerably increased in aged + M. pruriens. The diameter & volume of the seminiferous tubules, the height & volume of the epithelium, and the number of Leydig cells number were significantly decreased in aged rat testis, concomitantly connective tissue proportion was increased compared to adult rats. The seminiferous epithelium indicates significant rejuvenation or restoration of spermatogenic cells in aged + M. pruriens rat testis. The highlighting observations in aged + M. pruriens was increased in the following parameters i.e., tubular diameter (25%), number of tubules (35%), epithelial height (25%) & volume (20%), and number of Leydig cells (35%) when compared to untreated aged rat testis. The TNFα, NF-κB, cytochrome c, Caspase-9, Caspase-3, Bcl-2, Bax, PARP iNOS, and inflammatory and apoptotic factors were downregulated in aged + M. pruriens. M. pruriens was able to restore spermatogenesis and enhance the activity of Sertoli cells and Leydig cells and improve the pituitary-gonadal axis in aged rat testis and observations indicate the therapeutic activity of M. pruriens in aged rat testis.

Morphological diversity of sperm: A mini review.

Sperms are highly specialized cells for delivering DNA from male to the ovum. Incredibly, wide degree of diversity in sperm morphology in their basic structures i.e. head, middle piece and tail is found across species. Differences in terms of overall size of the sperm, shape and number of sperm produced are also incredible. One of the key for this variations or diversity in sperm may be associated with female reproductive tract, sperm competition, testicular size and sperm size and number. Establishing a correlation between sperm morphology and factors influencing them is a phenomenal task. In this mini-review these associations and the anatomical and functional adaptations among different from of sperm cells that have evolved to optimize fertilization success are discussed. Nevertheless, explaining these morphological diversities in sperm cells is a challenging question and it seems that evolutionary biologists have only recently engaged in exploring its links and patterns. From the literatures it seems that there is no causal relationship between sperm size and testicular size, however, the accumulated knowledge do indicates evolution of sperm morphology across species has some associations with female reproductive tract, sperm competition and sperm size and number, however interpreting these results for phylogentic correlations should be approached with caution.

Impact of sperm retrieval [corrected] on testis and epididymis: an experimental study using Wistar albino rats.

The aim of this study was to analyze pathophysiological changes after testicular sperm aspiration (TESA) and microsurgical epididymal sperm aspiration (MESA) procedures. Twenty four mature male Wistar albino rats with a proven breeding history, weighing approximately 200-250 gm were used for the study. Animals were randomly divided into four groups (n = 6), i.e., control, sham-control, unilateral TESA, and MESA. Using a 22G needle, the aspiration procedures were done in testis or caudal epididymis. At the end of 60 days of survival, blood samples were collected and processed for antisperm antibody detection by enzyme-linked immunosorbent assay (ELISA). After euthanasia, testes and epididymides were collected and processed for paraffin embedding. Sections were stained with hematoxylin and eosin, and TUNEL technique. Serum antisperm antibody titer significantly increased in TESA (P < 0.001) when compared to MESA. Histomorphometric analysis indicated testicular alterations in TESA and MESA, with significant damage in TESA in both testes (P < 0.001). Following the MESA procedure, ipsilateral caudal and carpus epididymis showed significant alterations (P < 0.001) and no such alterations were seen in the ipsilateral caput and intact contralateral epididymis. TUNEL staining revealed an up-regulation of apoptosis in both contra- and ipsilateral testes of TESA. Needle prick had produced drastic and irreversible alterations in testis of TESA. Ensuing processes of immunological and inflammatory reaction had the potential to disrupt spermatogenesis and increase germ cell apoptosis. However, extrapolating conclusions from the experimental model to the clinic needs to be done cautiously.

Effect of Mucuna pruriens (Linn.) on mitochondrial dysfunction and DNA damage in epididymal sperm of streptozotocin induced diabetic rat.

ETHNOPHARMACOLOGICAL RELEVANCE: Mucuna pruriens Linn. (M. pruriens) is a leguminous plant that has been recognized as an herbal medicine for improving fertility and related disorders in the Indian traditional system of medicine, however without proper scientific validations. AIM OF THE STUDY: To study the effect of ethanolic seed extract of M. pruriens on mitochondrial dysfunction and the DNA damage in hyperglycemic rat epididymal spermatozoa. MATERIALS AND METHODS: Male Wistar albino rats were divided as control (Sham), diabetes induced [streptozotocin 60 mg/kg of body weight (b.w.) in 0.1M citrate buffer] (STZ), diabetic rats administered with 200mg/kg b.w. of extract (STZ+MP) and normal rats administered with 200mg/kg b.w. of extract (Sham+MP). M. pruriens was administered (gavage) once daily for a period of 60 days. On 60th day animals were sacrificed by cervical dislocation sperm were collected from epididymis and subjected various analysis like antioxidants, ROS, lipid peroxidation (LPO), DNA damage, chromosomal integrity and mitochondrial membrane potential (MMP). RESULTS: Significant reduction in the sperm count, motility, viability and significant increase in the number of abnormal sperm in STZ compared to sham was noticed. STZ rat sperm showed significant increase in LPO and DNA damage. Both the enzymic and non-enzymic were decreased; MMP and the mitochondrial functions were severely affected in STZ group. The diabetic rats supplemented with M. pruriens showed a remarkable recovery in antioxidant levels and reduced LPO with well preserved sperm DNA. MMP and mitochondrial function test were also preserved in STZ+MP rat sperm. CONCLUSION: The present study has clearly demonstrated the potency of M. pruriens to reduce the diabetic induced sperm damage induced by oxidative stress (OS). These observations are encouraging to perform similar studies in human.

Morphological analysis of the maxillary arch and hard palate in edentulous maxilla of South Indian dry skulls.

PURPOSE: The purpose of this study is to demonstrate the role of aging\edentulousness on the maxillary arch, the size of the alveolar process, the shape and thickness of the hard palate in the South Indian dry skulls to customize more appropriate treatment of elderly edentulous patients. METHODS: One hundred dry skulls were divided into dentate and edentulous groups and were subgrouped into male and female. They were subjected to various morphological and morphometrical analyses. RESULTS: The data have revealed a more significant reduction in the depth and width (p < 0.001) of the arch corresponding to the reduction in the size and thickness of the alveolar process (p < 0.001) in the edentulous skull. Further, a significant reduction in the size and thickness (p < 0.001) of the hard palate was observed in the edentulous skull. The data clearly indicate the effect of tooth loss in the horizontal regression of the maxilla. Furthermore, it may enhance the bone resorption that can ultimately result in the marked reduction of the width and depth of the alveolar arch concomitant with the alteration of the hard palate. These changes may subsequently alter the facial appearance in the elderly population. CONCLUSION: The observed data from specific group of population may provide relevant data for their comparative analysis between different populations for a better understanding of their regional differences with respect to environmental and social influence. Moreover, the data can provide a better idea to evaluate a promising treatment strategy in prosthodontics and orthodontics in South India.

Effect of Mucuna pruriens (Linn.) on sexual behavior and sperm parameters in streptozotocin-induced diabetic male rat.

INTRODUCTION: Sexual dysfunction is one of the major secondary complications in the diabetic. Mucuna pruriens, a leguminous plant identified for its antidiabetic, aphrodisiac, and improving fertility properties, has been the choice of Indian traditional medicine. AIM: Objective of the present study was to analyze the efficacy of M. pruriens on male sexual behavior and sperm parameters in long-term hyperglycemic male rats. METHODS: Male albino rats were divided as group I control, group II diabetes induced (streptozotocin [STZ] 60 mg/kg of body weight (b.w.) in 0.1 M citrate buffer), group III diabetic rats administered with 200 mg/kg b.w. of ethanolic extract of M. pruriens seed, group IV diabetic rats administered with 5 mg/kg b.w. of sildenafil citrate (SC), group V administered with 200 mg/kg b.w. of extract, and group VI administered with 5 mg/kg b.w. of SC. M. pruriens and SC were administered in single oral dosage per day for a period of 60 days. The animals were subjected to mating behavior analyses, libido, test of potency, and epididymal sperms were analyzed. MAIN OUTCOME MEASURE: The mating behavior, libido, test of potency, along with epididymal sperms were studied. RESULTS: The study showed significant reduction in sexual behavior and sperm parameters in group II. Daily sperm production (DSP) and levels of follicular stimulating hormone, luteinizing hormone, and testosterone were significantly reduced in group II, whereas the animals with diabetes administered with seed extract of M. pruriens (group III) showed significant improvement in sexual behavior, libido and potency, sperm parameters, DSP, and hormonal levels when compared to group II. CONCLUSION: The present work reveals the potential efficacy of ethanolic seed extract of M. pruriens to improve male sexual behavior with androgenic and antidiabetic effects in the STZ-induced diabetic male rats. This study supports the usage of M. pruriens in the Indian system of medicine as sexual invigorator in diabetic condition and encourages performing similar study in men.

Effect of Mucuna pruriens (Linn.) on oxidative stress-induced structural alteration of corpus cavernosum in streptozotocin-induced diabetic rat.

INTRODUCTION: Erectile dysfunction is one of the major secondary complications of diabetes. Mucuna pruriens (M. pruriens), a leguminous plant identified for its antidiabetic, aphrodisiac, and fertility enhancing properties, has been the choice of Indian traditional medicine. AIM: The objective of the present study was to analyze the efficacy of M. pruriens on free radicals-mediated penile tissue alterations in hyperglycemic male rats. Methods. Male albino rats were divided as group I (sham) control, group II (STZ) diabetes-induced (streptozotocin 60 mg/kg of body weight [bw] in 0.1 M citrate buffer), group III (STZ + MP) diabetic rats administered with 200 mg/kg bw of ethanolic extract of M. pruriens seed, group IV (STZ + SIL) diabetic rats administered with 5 mg/kg bw of sildenafil citrate, group V (sham + MP) administered with 200 mg/kg bw of extract alone, and group VI (sham + SIL) administered with 5 mg/kg bw of sildenafil citrate. The M. pruriens and sildenafil citrate were given (gavage) once daily for a period of 60 days. At the end of 60 days, the animals were sacrificed and subjected to analysis of reactive oxygen species levels, enzymic and nonenzymic antioxidant levels, levels of NOx, histological, and histomorphometrical study of penile tissue. MAIN OUTCOME MEASURES: Remedial use of M. pruriens seed extract on diabetes-induced erectile tissue damage. RESULTS: Significantly high levels of oxidative stress and low levels of antioxidants in the penile tissue seem to contribute to the increased collagen deposition and fibrosis of erectile tissue in STZ rats. Relatively, there was increased damage in STZ + SIL group. Supplementation of M. pruriens in STZ + MP group has revealed the potency to overcome oxidative stress, and good preservation of penile histoarchitecture. CONCLUSION: The ethanolic extract of M. pruriens seed significantly recovered or protected erectile tissue from the oxidative stress-induced degeneration by its antioxidant potentials. These findings propound to serve mankind by the treatment of diabetes-induced erectile dysfunction.

Red clover Trifolium pratense (Linn.) isoflavones extract on the pain threshold of normal and ovariectomized rats--a long-term study.

Depletion of estrogens occurs in women during menopause, while in experimental animals, oophorectomy is a common method to deplete the animals of their gonadal hormones. Recently, phytoestrogens derived from plants have been tried as estrogen substitutes during menopause. In the present study an isoflavones methanol extract from red clover Trifolium pratense (Linn.) was administered orally (500 mg/kg of body weight) to ovariectomized (OVX) and normal (controls) rats for 90 and 180 days. Their pain threshold was monitored using tail flicking and formalin test methods. Observations showed that the OVX rat pain threshold was reduced due to estrogen deprivation, whereas the pain threshold levels in OVX rats treated with isoflavones extract was similar to the control animals. The present study demonstrated the influence of phytoestrogen on long-term OVX rats in pain perception in the absence of ovarian estrogen and without toxic side effects. However, the actions of gonadal hormones on nociceptive axis are myriad and complex, so further studies on the exact physiological mechanism of the phytoestrogen action on nociceptive axis is warranted.

Effect of Mucuna pruriens on oxidative stress mediated damage in aged rat sperm.

Mucuna pruriens Linn., a leguminous plant, has been recognized as an aphrodisiac and spermatogenic agent. Protective efficacy of M. pruriens on reactive oxygen species (ROS)-induced pathophysiological alterations in structural and functional integrity of epididymal sperm in aged Wister albino rat was analysed. Animals were grouped as groups I, II, III and IV, i.e. young (control), aged, aged treated with ethanolic extract (200 mg/kg b.w.) of M. pruriens and young rats treated with M. pruriens, respectively. At the end of the experimental period, i.e. after 60 days animals were sacrificed, epididymal sperm were collected and subjected to count, viability, motility, morphology and morphometric analysis. Enzymatic and non-enzymatic antioxidants, ROS, lipid peroxidation (LPO), DNA damage, chromosomal integrity and mitochondrial membrane potential were estimated. Results obtained from the aged animals showed significant reduction in sperm count, viability and motility, increased morphological damage and an increase in the number of sperm with cytoplasmic remnant, and these alterations were significantly reversed in M. pruriens treated group. Significant increase in LPO, HO and H(2)O(2) production and significant decline in the levels of the enzymatic and non-enzymatic antioxidants were observed in the aged animals. Supplementation of M. pruriens significantly reduced ROS and LPO production and significant increase in both enzymatic and non-enzymatic antioxidant levels. There were significant DNA damage, loss of chromosomal integrity and increase in mitochondrial membrane permeability in aged rat sperm. This was significantly reduced in group III. Present observation indicates the antioxidant enhancing property, free radical quenching ability and spermatogenic efficacy of the M. pruriens. Collectively, sperm damage in ageing was significantly reduced by quenching ROS, improving antioxidant defence system and mitochondrial function.

Withania somnifera root extract improves catecholamines and physiological abnormalities seen in a Parkinson's disease model mouse.

ETHNOPHARMACOLOGICAL RELEVANCE: Withania somnifera root extract (Ws)/Ashwagandha/Indian ginseng is a traditional herbal medicine, used over 4000 years in India, shown to have effect on neural growth and locomotor function. Although catecholamines and oxidative stress resulting in neurodegeneration and locomotor disorder are the main events in Parkinson's disease (PD), efficacy of the drug on these molecules and physiological abnormality are not clear. AIM OF THE STUDY: The objective of the study was to examine effect of Ws on catecholamines and physiological abnormalities seen in PD using PD model mouse. MATERIALS AND METHODS: Mouse were treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 4 days to show biochemical and physiological abnormalities similar to patients with PD. PD mice were treated with Ws 100mg/kg body weight for 7 or 28 days. Catecholamines: dopamine (DA), 3,4-dihydroxy-phenylacetic acid (DOPAC) and homovanillic acid (HVA); antioxidants: glutathione (GSH) and glutathione peroxidase (GPx); and lipid peroxidation marker (TBARS) were analyzed in the Ws treated and untreated PD mouse striatum. RESULTS: Mouse treated with MPTP showed reduced levels of DA, DOPAC, HVA, GSH and GPx and induced thiobarbituric acid reactive substance (TBARS) level compared to the control. Physiological abnormalities were seen in the mouse as determined by hang test and rotarod test. Oral treatment of PD mouse Ws root extract (100mg/kg body weight) for 7 days or 28 days increased DA, DOPAC and HVA levels and normalized TBARS levels in the corpus striatum of the PD mouse. The 7 days Ws treated mice showed improved motor function as determined by hang test and rotarod test. Treatment with Ws for 28 days increased GSH and GPx levels in the striatum compared to the Ws untreated PD mouse striatum. CONCLUSION: These data suggest that Ws is a potential drug in treating catecholamines, oxidative damage and physiological abnormalities seen in the PD mouse.

Dose- and time-dependent effects of ethanolic extract of Mucuna pruriens Linn. seed on sexual behaviour of normal male rats.

AIM OF THE STUDY: According to Indian Systems of Medicine, Mucuna pruriens Linn., belonging to the leguminous family (Papilionaceae), were used for treating male sexual disorders since ancient times. In this study, the effects of ethanolic extracts of the Mucuna pruriens Linn. seed on general mating behaviour, libido and potency of normal male Wister albino rats were investigated and also compared with the standard reference drug, Sildenafil citrate. MATERIALS AND METHODS: Animals were divided into one control group (Group I--received saline) and four experimental groups (Groups II-V). Experimental groups were divided on the basis of the dosage of extract to the animals as follows: 150 mg/kg body weight (Group I), 200mg/kg body weight (Group II) and 250 mg/kg body weight (Group IV) while Group V received Sildenafil citrate (5mg/kg body weight). Animals were fed PO with saline or extract or standard drug once in a day for 45 days. To analyse the mating behaviour, female rats with oestrus phase were used. RESULTS: The extract administered PO significantly increased the mounting frequency, intromission frequency and ejaculation latency, and decreased the mounting latency, intromission latency, post-ejaculatory interval and inter-intromission interval. The potency test significantly increased erections, quick flips, long flips and total reflex. Therefore, the results indicated that the ethanolic extracts of Mucuna pruriens Linn. seed produced a significant and sustained increase in the sexual activity of normal male rats at a particular dose (200mg/kg). When compared to control, all the drug-treated groups have shown drug-induced effects for a few parameters. However in Group II, there was an obvious enhancement in all parameters, without affecting the normal behaviour. When compared with the standard drug, the net effect of extract is even less than that in Group II. CONCLUSIONS: Therefore, the resulting aphrodisiac activity of the extract lends support to the claim that it has traditionally been used for the treatment of sexual disorders.

Dracunculiasis in South Indian bonnet monkey.

Dracunculiasis, popularly known as Guinea worm disease, has been eradicated from Tamil Nadu, India, and there have been no indigenous cases reported since 1981. This report describes a female bonnet monkey with dracunculiasis. She presented with fever and a blister in left hind limb. The blister ruptured on exposure to water and a 7-cm-long worm was extruded. The worm died before it could be histologically examined. The diagnosis was based on the typical clinical course, which was pathognomonic of dracunculiasis. Review of literature did not reveal any previous report of dracunculiasis in South Indian bonnet monkeys (Macaca radiata). This paper raises the question whether wild monkeys might act as reservoirs of human infection and cause resurgence of the disease in South India.